Abstract
Accumulation of alpha-synuclein (α-syn) is central to the pathogenesis of Parkinson’s disease (PD). Previous studies suggest that α-syn pathology may originate from the olfactory bulb (OB) or gut in response to an unknown pathogen and later progress to the different brain regions. Aging is viewed as the utmost threat to PD development. Therefore, studies depicting the role of age in α-syn accumulation and its progression in PD are important. In the present study, we gave intranasal rotenone microemulsion for 6 weeks in 12-month-old female BALB/c mice and found olfactory dysfunction after 4 and 6 weeks of rotenone administration. Interestingly, motor impairment was observed only after 6 weeks. The animals were sacrificed after 6 weeks to perform western blotting and immunohistochemical studies to detect α-syn pathology, neuroinflammation and neurodegeneration. We found α-syn accumulation in OB, striatum, substantia nigra (SN) and cortex. Importantly, we found significant glial cell activation and neurodegeneration in all the analysed regions which were absent in our previous published studies with 3 months old mice even after they were exposed to rotenone for 9 weeks indicating age is a crucial factor for α-syn induced neuroinflammation and neurodegeneration. We also observed increased iron accumulation in SN of rotenone-exposed aged mice. Moreover, inflammaging was observed in OB and striatum of 12-month-old BALB/c mice as compared to 3-month-old BALB/c mice. In conclusion, there is a difference in sensitivity between adult and aged mice in the development and progression of α-syn pathology and subsequent neurodegeneration, for which inflammaging might be the crucial probable mechanism.
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Data Availability
The datasets used during the present study are available from the corresponding author upon reasonable request.
Abbreviations
- α-syn:
-
Alpha-synuclein
- BCA:
-
Bicinchoninic acid
- CPCSEA:
-
Committee for the Purpose of Control and Supervision of Experiments on animals
- DAB:
-
3,3′-Diaminobenzidine
- DA:
-
Dopamine
- DAergic:
-
Dopaminergic
- DPX:
-
Dibutylphthalate Polystyrene Xylene
- CNS:
-
Central nervous system
- GFAP:
-
Glial fibrillary acidic protein
- IAEC:
-
Institutional animal ethics committee
- Iba-1:
-
Ionized calcium binding adaptor molecule 1
- IHC:
-
Immunohistochemistry
- i.n:
-
Intranasal
- mAb:
-
Monoclonal antibody
- ME:
-
Microemulsion
- NFkB:
-
Nuclear Factor Kappa B
- OB:
-
Olfactory bulb
- pAb:
-
Polyclonal antibody
- PBS:
-
Phosphate buffer saline
- PD:
-
Parkinson’s disease
- PFA:
-
Paraformaldehyde
- PMSF:
-
Phenylmethyl sulfonyl fluoride
- pTH:
-
Phosphorylated tyrosine hydroxylase
- psyn:
-
Phosphorylated alpha-synuclein
- PVDF:
-
Polyvinylidene fluoride
- RIPA buffer:
-
Radioimmunoprecipitation assay buffer
- RBWT:
-
Round beam walk test
- SDS:
-
Sodium dodecyl sulphate
- SLS:
-
Sodium lauryl sulphate
- SN:
-
Substantia nigra
- SNc:
-
Substantia nigra pars compacta
- TEMED:
-
Tetramethyl-ethylenediamine
- TH:
-
Tyrosine hydroxylase
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Acknowledgements
The authors acknowledge National Institute of Pharmaceutical Education and Research (NIPER) Ahmedabad administration for providing the facility and support for conducting this study. They are supported by project no. LX22NPO5107 (MEYS): financed by European Union – Next Generation EU.
Funding
This supplement was supported by the National Institute of Pharmaceutical Education and Research (NIPER) seed fund-Ahmedabad, Department of Pharmaceutics, Ministry of Chemicals and Fertilizers, Government of India. Dr. Amit Khairnar gratefully acknowledges the support of the Ramalingaswami Fellowship (No. BT/RLF/Re-entry/24/2017) from the Department of Biotechnology, India.
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Conception and study design was done by AK and MS. Data acquisition was done by MS and NS. Data was analysed by MS and NS. Manuscript was written by MS and revised by AK, MS and NS.
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No clinical study was conducted. Experimental design for pre-clinical study was approved by IAEC Committee of NIPER Ahmedabad (IAEC approval number: NIPERA/IAEC/2019/19). All efforts were made to minimize animal suffering and to reduce the number of animals used.
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Sharma, M., Sharma, N. & Khairnar, A. Intranasal Rotenone Induces Alpha-Synuclein Accumulation, Neuroinflammation and Dopaminergic Neurodegeneration in Middle-Aged Mice. Neurochem Res 48, 1543–1560 (2023). https://doi.org/10.1007/s11064-022-03847-y
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DOI: https://doi.org/10.1007/s11064-022-03847-y